[The basic properties of mesenchymal stromal cells from the donor bone marrow: superficial markers].

AIM: To characterize a superficial phenotype and to make a cytogenetic analysis of bone marrow (BM) mesenchymal stromal cells (MSC) from donors. MATERIALS AND METHODS: The study analyzed BM samples from 11 healthy donors. The phenotype of obtained MSC was analyzed using cytofluorometry. Chromosomal analysis was carried out at the first-second passage. RESULTS: The superficial phenotype of MSC was steady-state during 8 passages and conformed to the worldwide standard for this cell population. The marker NGFR+ was detectable only during the first 2 passages and the count of CD146+ cells was decreased to 50% as consecutive passages were carried out, which confirms that MSCs have lost their neural and endothelial differentiation capacity. MSCs are stably able to differentiate only into the mesenchymal lineage. The detection of chromosomal rearrangements in MSCs at different stages of cultivation revealed no clonal rearrangements in any case. However, chromosomal aberrations were found 3-10% of metaphases at the first and second passages, which may be associated with chromosome instability in primary cultures. CONCLUSION: The pooled data suggest that the analyzed MSCs meet the conventional worldwide standards.

[Efficiency of cyclosporin A therapy in patients with myelodysplastic syndrome].

AIM: To evaluate the efficacy of cyclosporin A (CsA) in patients with myelodysplastic syndromes (MDS) and to identify determinants of a response to this therapy. SUBJECTS AND METHODS: The efficacy of CsA was evaluated in 52 patients (30 men and 22 women aged 16 to 74 years) with MDS. Thirty-two patients were given CsA as first-line therapy; 20 patients took the agent after prior therapy. CsA was used in a daily oral dose of 5 mg/kg. Its efficacy was evaluated following 3, 6, and 12 months. Actuarial survival was determined by the Kaplan-Meier method. RESULTS: The efficacy of CsA used as first- and second-line therapy was 56 and 55%, respectively; complete remissions were achieved in 19 and 20% of cases. Baseline refractory anemia (RA) transformed to RA with excess blasts (RAEB) in 31% of cases; baseline RAEB did to acute myeloid leukemia in 34%. Overall survival was significantly associated with bone marrow (BM) blast cell percentage (< 5% or > 5%; p = 0.0009), BM cellularity (hypoplasia and focal hypoplasia of hematopoiesis or BM hyperplasia; p = 0.03), focal polyclonal lymphoid infiltration in the BM (p = 0.01) and karyotype anomalies (low, moderate, and high risks; p = 0.001). CONCLUSION: CsA is the drug of choice in treating patients with MDS, including RA, RA with ringed sideroblasts, refractory cytopenia with multilineage dysplasia, with hypoplasia of hematopoiesis, with nodular polyclonal lymphoid infiltration in the BM, a normal karyotype or changes corresponding to a low or moderate IPSS risk.

[Allogenic transplantation of hemopoietic stem cells in low-intensity regimes in patients with hematological malignancies].

AIM: To test feasibility of transplantation of hemopoietic stem cells (THSC) with conditioning in low-intensity regimen associated with minimal toxic complications and engraftment in patients with hematological malignancy (HM) from a high risk group. MATERIAL AND METHODS: THSC was performed in 33 patients aged 18 to 65 years. Most of the patients suffered from acute leukemia and advanced forms of myelodysplastic syndrome. All the patients had severe complications excluding standard transplantation. Pretransplantation preparation was based on fludarabine and moderate doses of busulfane. Engraftment was achieved in 94% patients. Of complications, there were primarily infections, relapses, graft versus host reactions (45, 24, 57.5%, respectively). Overall survival was 53%, relapse-free--67%, follow-up median 23.6 months. CONCLUSION: THSC after conditioning in the regime of low intensity is an effective method of HM treatment in patients with contraindications to standard transplantation. The main problem is a high risk to develop graft versus host reaction, especially a chronic form.

[Monitoring of minimal residual disease in patients with acute promyelocytic leukemia].

AIM: To study efficacy of different programs of maintenance therapy, to create the program of differential therapy of minimal residual disease (MRD) and molecular recurrences at all stages of acute promyelocytic leukemia (APL) basing on the results of monitoring. MATERIAL AND METHODS: A total of 76 APL patients entered the trial. They received therapy by the protocols APL-97/98, APL-01, AIDA, 5D. Expression of chymeric oncogen PML/RARa in the disease onset was estimated by bone marrow and/or peripheral blood examination with RT-PCR. The study of chymeric oncogen PML/RARa was made once in two months. RESULTS: The program of differential therapy of APL is proposed on the basis of molecular-biological monitoring of expression of chymeric oncogen PML/RARa. The results of molecular monitoring of MRD correlated with development of molecular and hematological recurrences. Therapeutic policy is determined after diagnosis of molecular recurrence. Further therapy of APL is determined which allows a rise in overall and recurrence-free survival of the patients. CONCLUSION: The efficacy of maintenance therapy only with cytostatic drugs or their combination with ATRA is similar. The response to biological therapy with ATRA plus interferon-alpha is not sufficient. Molecular recurrences--probable or documented--are detected in maintenance therapy 2 months earlier, on the average, than hematological ones. Changes in the treatment policy in registration of molecular recurrence significantly diminish probability of hematological recurrence (from 36 to 0%, p = 0.001.

[Clonal chromosomal aberrations in patients with aplastic anemia at the disease onset and transformation].

AIM: To estimate detectability and characteristic features of chromosomal aberrations in bone marrow cells of patients with aplastic anemia (AA). MATERIAL AND METHODS: The trial covered 155 AA patients admitted to the Hematological Research Center in 1987-2002. Cytogenetic study by G-differential staining was performed in 58 patients with AA and 5 patients with AA transforming into myelodysplastic syndrome (MDS) or acute leukemia (AL). Cytogenetic and morphological specimens of the latter's bone marrow were studied retrospectively using fluorescent in situ hybridization (FISH) with DNA probes for detection of monosomia 7 and deletion 7q. RESULTS: Clonal chromosomal aberrations were detected in 4 out of 28 patients. Further examinations revealed no aberrations. Clonal diseases developed in 7 (4.5%) of 155 patients. In 2 patients the disease transformed into paroxysmal nocturnal hemoglobinuria, 5 (3.2%) patients developed variants of MDS and AL. Monosomia 7 or deletion 7q were diagnosed in 3 cases of MDS/AL. In retrospective study of bone marrow specimens of patients with transformation in MDS/AL with monosomia 7, FISH recognized a small elevation over control values in 2 cases. CONCLUSION: Stable clonal chromosomal aberrations are not characteristic of AA. Some AA patients with subsequent MDS/AL may have minor neoplastic clone in the disease onset.

[Chromosomal aberrations in myelodysplastic syndrome].

AIM: To analyse incidence rate of chromosomal aberrations in myelodysplastic syndromes (MDS), specification of clinicomorphological features of some cytogenetic variants. MATERIAL AND METHODS: Chromosomal analysis by the method of G-differential staining of chromosomes was made in 209 patients with different variants of MDS. RESULTS; Clonal chromosomal aberrations occured in 60.8%. The following aberrations were found most frequently: deletion of the long arm of the chromosome 5 (del(5q)) - 34.6%, trisomy of chromosome 8 (14.1%), monosomy of chromosome 7 (13.4%), aberrations 3q21q26 (12.6%), aberrations of a long arm of X-chromosome (4.7%), the absence of Y-chromosome (3.1%). Complex aberrations of karyotype were found in 13.5% cases. Chromosomal aberrations determined not only clinical and morphological features but also the prognosis of the disease. CONCLUSION: Cytogenetic examination is an essential component of MDS patients examination. It allows more precise classification of MDS variant and prognostification of the disease course.

[Preliminary results of a multicenter randomized study on the treatment of acute promyelocytic leukemias]. / Predvaritel'nye rezul'taty mnogotsentrovogo randomizirovannogo issledovaniia po lecheniiu ostrykh promielotsitarnykh leikozov.

AIM: To study efficacy of maintenance therapy of patients with acute promyelocytic leukemia (APL) in the APL treatment Russian multicenter trial. MATERIAL AND METHODS: The trial was made with participation of 18 hematological departments of clinics in Russia. A total of 68 APL patients entered the trial. The maintenance therapy consisted of 5-day courses of cytostatic drugs which alternated or did not alternate with 5-day courses of ATRA. Cytogenetic tests were made in 31 patients, t(15;17) was detected in 26 of them. Molecular examination conducted in 28 patients discovered chimeric transcript PML/RARa in 26 of them. Of 20 patients examined in Hematological Research Center, 7 (35%) had a bcr 1/2 variant of the transcript PML/RARa, 13 (65%)--bcr 3 variant. RESULTS: 65 patients were eligible for assessment. A complete remission was achieved in 90% cases. No resistance was observed. In follow-up within 30 months the recurrence rate was similar on both treatments. The results of the induction therapy and survival in patients with different variants of the transcripts were also similar. Overall 2.5 year survival for all the patients was 77%, recurrence-free--80%. The survival analysis in patients with leukocytosis higher and lower 10 x 10(9)/l found no statistical differences by the survival. Patients with hyperleukocytosis had higher early lethality than patients with leukocytes under 10 x 10(9)/l (25% vs 5.3%, p = 0.03). CONCLUSION: The APL 06.01 protocol showed high efficacy of the relevant maintenance which provides a complete molecular remission in the majority of patients with probable recurrence-free 2.5 year survival 80%.

[Allogenic bone marrow transplantation after reduced intensity conditioning regimens in therapy of patients with hemoblastoses]. / Allogennaia transplantatsiia kostnogo mozga posle rezhimov konditsionirovaniia ponizhennoi intensivnosti v terapii bol'nykh gemoblastozami.

AIM: To investigate effectiveness of allogenic transplantation of the bone marrow (TBM) in the treatment of hemoblastosis patients from a high risk group, the course of donor bone marrow retention, tolerance and antitumor activity of this therapy. MATERIAL AND METHODS: 11 patients received TBM in low-intensity regimen in Hematological Research Center in 1999-2001. All the patients were from a high risk group. Conditioning was based on the combination of fludarabin with busulfan. The transplanted precursor cells were taken from the bone marrow and/or peripheral donor blood. The retention was controlled by differential agglutination of erythrocytes and amplification of hypervariable sites of DNA. Minimal residual disease was controlled by standard cytogenetical tests, fluorescent in situ hybridization or reverse-transcriptase polymerase chain reaction. RESULTS: All the patients tolerated pretransplantation conditioning well. By chimerism, signs of retention of donor bone marrow on day +30 after TBM were observed in 9 patients of 11. Acute graft versus host reaction developed in 5 patients. This reaction was treated conventionally with methylprednisone and cyclosporin A, in 4 cases with a good effect. A complete remission persists in 5 patients. Mean follow-up lasted for 241 days. CONCLUSION: Thus, transplantation was successful in 50% patients with an unfavourable prognosis who are still in a complete remission. This suggests efficacy of the above method of treatment.

[Liposomal daunorubicin and interferon-alfa in combination with all-retinoic acid--a new regime for the treatment of acute promyelocytic leukemia]. / Liposomal'nyi daunorubitsin i interfron-al'fa v sochetanii s polnost'iu transretinoevoi kislotoi--novaia programma lecheniia ostrykh promielotsitarnykh leikozov.

AIM: To define duration and schemes of maintenance therapy, to evaluate cytarabine demand while treatment of acute promyelocytic leukemia (APL) with an original program 5D + Ifa + AT RA. MATERIAL AND METHODS: The above treatment was conducted in 7 patients with APL (1 male, 6 females, 18-45 years, leukocytes 1.2-15.0 x 10(9)/l). RESULTS: Induction by 5D program was performed in 5 patients. A complete remission was achieved in 4 of them, molecular--in 3. One woman died. 5D consolidation was performed in 6 patients. After the first course of consolidation all the patients achieved molecular remission. Maintenance of remission with Ifa + ATRA was made in 5 patients. In one female the remission came to the end. Mean time of the follow-up was 18 months. The remission lasted 5-36 months. Monitoring of molecular remission has not found marker PML/RARa. CONCLUSION: Daunozom monotherapy leads to a complete remission after the first course in most of the patients. In the majority of them it was molecular. Maintenance with If + ATRA for 2 years maintains molecular remission for 5-36 months.

[Molecular cytogenetic monitoring of chimerism and minimal residual disease in patient with chronic myeloid leukemia after allogenic and syngenic bone marrow transplantation]. / Molekuliarno-tsitogeneticheskii monitoring khimerizma i minimal'noi ostatochnoi bolezni u bol'nykh khronicheskim mieloleikozom posle allogennoi i singennoi transplantatsii kostnogo mozga.

AIM: To study trends in restoration of normal and tumor hemopoiesis after transplantation of allogenic and syngenic hemopoietic cells. MATERIAL AND METHODS: The examination of bone marrow before transplantation and 1, 2, 3, 6, 9 months, 1, 2 and 3 years after bone marrow transplantation (BMT) was made in 25 patients with chronic myeloid leukemia (CML) after allogenic transplantation of the bone marrow (TBM) and 4 patients after syngenic TBM. The method of G-differential staining of chromosomes and fluorescent hybridization in situ (FISH) with DNA probe to centromeric sites X/Y of chromosomes and genes BCR/ABL was used. RESULTS: 56% of CML patients after allogenic BMT were in cytogenetic and clinicohematological remission, 16% developed early cytogenetic recurrence. Single metaphase with t(9;22) were identified in 28%; 14.3% developed late cytogenetic and hematological recurrence. In patients in posttransplantation remission there were from 0.1 to 5.8% cells of the host. The number of cells of the host and the number of BCR/ABL-positive cells correlated significantly. CONCLUSION: The results of 8-year monitoring of chimerism and minimal residual disease validate application of molecular-cytogenetic methods for assessing transplant condition.

[Induced leukemias and their connection with radiation exposure]. / Indutsirovannye leikozy, ikh sviaz' s vozdeistviem radiatsii.

The cytogenetic study of bone marrow cells was performed in 40 patients with secondary leukemias which have arisen after application of cytostatic and/or radiotherapy for primary tumours (Hodgkin's disease, lymphomas, acute lymphoblastic leukemias, breast cancer and other solid tumours). The comparative analysis of results has shown, that the leukemias after irradiating or application of alkylating agents and irradiation, have the quite particular clinico-morphological and cytogenetic characteristics. In 70% of cases these diseases develops as smouldering leukemias with subsequent transformation in M-4, M-6, and rarely M-2 cytochemical variants. Primary cytogenetic events in 60% of researched karyotypes are the losses of long arms or whole chromosomes 5 and 7. In 20% of the researched cases normal karyotype was found, in the left 20%, the changes of a karyotype not including anomalies 5 and 7 chromosomes were detected. The obtained outcomes allow to consider the discharged complex of tags as reference for leukemias, induced by irradiating or chemical agents with similar mechanism of action (alkylating agents, benzene and its derivates). This complex of tags is typical for induced leukemias, and in a combination with definition of a level of stable aberrations in peripheral blood lymphocytes, can be utilised for abjection of radiation-induced leukemias from common mass of cases detected in regions, polluted by radionuclides. In this study in 60% of cases only specific for secondary leukemias chromosomal aberrations, including monosomy 5 and 7, rearrangements of 11q23 were found. On the base of the obtained data the differences in concepts of "secondary" and "induced" leukemias are considered.

[Combination of interferon-alpha and all-trans-retinoic acid as a treatment for maintaining remission in high-risk group patients with acute myeloid leukemia]. / Sochetanie interferona-alpha i polnost'iu trans-retinoevoi kisloty v kachestve programmy terapii podderzhaniia remissii u bol'nykh ostrymi mieloidnymi leikozami iz gruppy vysokogo riska.

AIM: To evaluate the efficiency of combination of interferon-alpha (INF) and all trans-retinoic acid (ATRA) as a treatment for maintaining remission in high risk group patients with acute myeloid leukemia (AML). MATERIALS AND METHODS: Three-day INF + ATRA course was administered every 3 months to 22 patients with AML from high risk group (impossibility of drug therapy during the first complete remission, resistant forms of AML, relapses, secondary AML, acute promyelocytic leukemia after attaining molecular remission). RESULTS: INF + ATRA during remission maintained a long first complete remission (median 18 months) in patients with primary AML after small-volume drug therapy, led to long first and second complete remissions (median 12 months) in patients with resistant AML, and induced and maintained molecular remissions in patients with acute promyelocytic leukemia.

[Results of clinical trials in the treatment of acute myeloid leukemia in adults during a 7-year period]. / Rezul'taty provodimykh v techenie 7 let klinicheskikh issledovanii po lecheniiu ostrykh mieloidnykh leikozov vzroslykh.

AIM: Comparison of effectiveness of induction regimens varying in intensity and maintenance variants in patients with acute myeloid leukemia (AML) included in a randomised multicenter trial. MATERIALS AND METHODS: Clinical trial 1 enrolled 185 AML patients. Vepesid-free induction was used in 85 patients (group 1), induction with vepesid--in 99 patients (group 2). 223 AML patients entered trial 2. Of them 37 patients were treated for 3 years 7 + 3 with daunorubicin in the dose 45 mg/m2 in the induction phase (group 1), 85 and 101 patients received 7 + 3 with daunorubicin for a year in the dose 45 mg/m2 and 60 mg/m2, respectively, (group 2 and 3). RESULTS: For group 1 in trial 1 the remission rate, early lethality, resistance were, respectively, 60, 20 and 20%, respectively. For group 2 in trial 1--66, 22 and 12%, respectively. 5-year recurrence-free survival reached 32 and 37% for group 1 and 2, respectively. For trial 2 relevant figures made up 75.5, 16.2, 8.1% for group 1; 60, 17.6 and 22.4% for group 2; 63, 20.8 and 16% for group 3, respectively. The 3.5-year recurrence-free survival in groups 1, 2 and 3, was 16, 46 and 50%, respectively. For both trials, the differences between the groups were insignificant. CONCLUSION: The results evidence that the treatment can be shorter (not 3 but 1 year or even 6 months), the induction more intensive (the dose of anthracycline antibiotics can be elevated up to 60 mg/m2 without a rise in early lethality).

[Effectiveness of trans-retinoic acid in the treatment of acute promyelocytic leukemia: initial results of a multicenter study]. / Effektivnost' polnost'iu trans-retinoevoi kisloty v terapii ostrykh promielotsitarnykh leikozov: pervye rezul'taty mnogotsentrovogo issledovaniia.

AIM: Evaluation of trans-retinoic acid (ATRA) in combination with cytostatic drugs in treatment of acute promyelocytic leukemia (APL). MATERIALS AND METHODS: In a multicenter study APL was treated according to protocols APL 01.97 and APL 06.87 in 28 patients (14 males and 14 females, median age 36 years). RESULTS: Administration of ATRA in combination with standard program 7 + 3 (cytosine-arabinoside 100 mg/m2 twice a day v.v. day 1-7, daunorubicin 60 mg/m2 v.v. day 1-3) induced a complete remission in 25 patients (90%). Early lethality was 10% (3 patients died). Resistant APL was not registered. Retinoid syndrome was diagnosed in 15 patients, one patient died. 2-year overall and recurrence-free survival made up 72 and 82%, respectively. CONCLUSION: ATRA combination with cytosine-arabinoside and daunorubicin is a novel treatment of acute promyelocytic leukemia providing a high rate of complete remission and long-term survival.